<italic toggle="yes">Pseudomonas aeruginosa</italic> AlgR Phosphorylation Status Differentially Regulates Pyocyanin and Pyoverdine Production

ABSTRACT Pseudomonas aeruginosa employs numerous, complex regulatory elements to control expression of its many virulence systems. The P. aeruginosa AlgZR two-component regulatory system controls the expression of several crucial virulence phenotypes. We recently determined, through transcriptomic p...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Alexander S. Little, Yuta Okkotsu, Alexandria A. Reinhart, F. Heath Damron, Mariette Barbier, Brandon Barrett, Amanda G. Oglesby-Sherrouse, Joanna B. Goldberg, William L. Cody, Michael J. Schurr, Michael L. Vasil
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	AlgR Pseudomonas aeruginosa iron acquisition pyocyanin pyoverdine virulence regulation Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/a049fc745130465a9bc5434bc9ba9c79
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:a049fc745130465a9bc5434bc9ba9c79
record_format	dspace
institution	DOAJ
collection	DOAJ
language	EN
topic	AlgR Pseudomonas aeruginosa iron acquisition pyocyanin pyoverdine virulence regulation Microbiology QR1-502
spellingShingle	AlgR Pseudomonas aeruginosa iron acquisition pyocyanin pyoverdine virulence regulation Microbiology QR1-502 Alexander S. Little Yuta Okkotsu Alexandria A. Reinhart F. Heath Damron Mariette Barbier Brandon Barrett Amanda G. Oglesby-Sherrouse Joanna B. Goldberg William L. Cody Michael J. Schurr Michael L. Vasil Michael J. Schurr <italic toggle="yes">Pseudomonas aeruginosa</italic> AlgR Phosphorylation Status Differentially Regulates Pyocyanin and Pyoverdine Production
description	ABSTRACT Pseudomonas aeruginosa employs numerous, complex regulatory elements to control expression of its many virulence systems. The P. aeruginosa AlgZR two-component regulatory system controls the expression of several crucial virulence phenotypes. We recently determined, through transcriptomic profiling of a PAO1 ΔalgR mutant strain compared to wild-type PAO1, that algZR and hemCD are cotranscribed and show differential iron-dependent gene expression. Previous expression profiling was performed in strains without algR and revealed that AlgR acts as either an activator or repressor, depending on the gene. Thus, examination of P. aeruginosa gene expression from cells locked into different AlgR phosphorylation states reveals greater physiological relevance. Therefore, gene expression from strains carrying algR alleles encoding a phosphomimetic (AlgR D54E) or a phosphoablative (AlgR D54N) form were compared by microarray to PAO1. Transcriptome analyses of these strains revealed 25 differentially expressed genes associated with iron siderophore biosynthesis or heme acquisition or production. The PAO1 algR D54N mutant produced lower levels of pyoverdine but increased expression of the small RNAs prrf1 and prrf2 compared to PAO1. In contrast, the algR D54N mutant produced more pyocyanin than wild-type PAO1. On the other hand, the PAO1 algR D54E mutant produced higher levels of pyoverdine, likely due to increased expression of an iron-regulated gene encoding the sigma factor pvdS, but it had decreased pyocyanin production. AlgR specifically bound to the prrf2 and pvdS promoters in vitro. AlgR-dependent pyoverdine production was additionally influenced by carbon source rather than the extracellular iron concentration per se. AlgR phosphorylation effects were also examined in a Drosophila melanogaster feeding, murine acute pneumonia, and punch wound infection models. Abrogation of AlgR phosphorylation attenuated P. aeruginosa virulence in these infection models. These results show that the AlgR phosphorylation state can directly, as well as indirectly, modulate the expression of iron acquisition genes that may ultimately impact the ability of P. aeruginosa to establish and maintain an infection. IMPORTANCE Pyoverdine and pyocyanin production are well-known P. aeruginosa virulence factors that obtain extracellular iron from the environment and from host proteins in different manners. Here, we show that the AlgR phosphorylation state inversely controls pyoverdine and pyocyanin production and that this control is carbon source dependent. P. aeruginosa expressing AlgR D54N, mimicking the constitutively unphosphorylated state, produced more pyocyanin than cells expressing wild-type AlgR. In contrast, a strain expressing an AlgR phosphomimetic (AlgR D54E) produced higher levels of pyoverdine. Pyoverdine production was directly controlled through the prrf2 small regulatory RNA and the pyoverdine sigma factor, PvdS. Abrogating pyoverdine or pyocyanin gene expression has been shown to attenuate virulence in a variety of models. Moreover, the inability to phosphorylate AlgR attenuates virulence in three different models, a Drosophila melanogaster feeding model, a murine acute pneumonia model, and a wound infection model. Interestingly, AlgR-dependent pyoverdine production was responsive to carbon source, indicating that this regulation has additional complexities that merit further study.
format	article
author	Alexander S. Little Yuta Okkotsu Alexandria A. Reinhart F. Heath Damron Mariette Barbier Brandon Barrett Amanda G. Oglesby-Sherrouse Joanna B. Goldberg William L. Cody Michael J. Schurr Michael L. Vasil Michael J. Schurr
author_facet	Alexander S. Little Yuta Okkotsu Alexandria A. Reinhart F. Heath Damron Mariette Barbier Brandon Barrett Amanda G. Oglesby-Sherrouse Joanna B. Goldberg William L. Cody Michael J. Schurr Michael L. Vasil Michael J. Schurr
author_sort	Alexander S. Little
title	<italic toggle="yes">Pseudomonas aeruginosa</italic> AlgR Phosphorylation Status Differentially Regulates Pyocyanin and Pyoverdine Production
title_short	<italic toggle="yes">Pseudomonas aeruginosa</italic> AlgR Phosphorylation Status Differentially Regulates Pyocyanin and Pyoverdine Production
title_full	<italic toggle="yes">Pseudomonas aeruginosa</italic> AlgR Phosphorylation Status Differentially Regulates Pyocyanin and Pyoverdine Production
title_fullStr	<italic toggle="yes">Pseudomonas aeruginosa</italic> AlgR Phosphorylation Status Differentially Regulates Pyocyanin and Pyoverdine Production
title_full_unstemmed	<italic toggle="yes">Pseudomonas aeruginosa</italic> AlgR Phosphorylation Status Differentially Regulates Pyocyanin and Pyoverdine Production
title_sort	<italic toggle="yes">pseudomonas aeruginosa</italic> algr phosphorylation status differentially regulates pyocyanin and pyoverdine production
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/a049fc745130465a9bc5434bc9ba9c79
work_keys_str_mv	AT alexanderslittle italictoggleyespseudomonasaeruginosaitalicalgrphosphorylationstatusdifferentiallyregulatespyocyaninandpyoverdineproduction AT yutaokkotsu italictoggleyespseudomonasaeruginosaitalicalgrphosphorylationstatusdifferentiallyregulatespyocyaninandpyoverdineproduction AT alexandriaareinhart italictoggleyespseudomonasaeruginosaitalicalgrphosphorylationstatusdifferentiallyregulatespyocyaninandpyoverdineproduction AT fheathdamron italictoggleyespseudomonasaeruginosaitalicalgrphosphorylationstatusdifferentiallyregulatespyocyaninandpyoverdineproduction AT mariettebarbier italictoggleyespseudomonasaeruginosaitalicalgrphosphorylationstatusdifferentiallyregulatespyocyaninandpyoverdineproduction AT brandonbarrett italictoggleyespseudomonasaeruginosaitalicalgrphosphorylationstatusdifferentiallyregulatespyocyaninandpyoverdineproduction AT amandagoglesbysherrouse italictoggleyespseudomonasaeruginosaitalicalgrphosphorylationstatusdifferentiallyregulatespyocyaninandpyoverdineproduction AT joannabgoldberg italictoggleyespseudomonasaeruginosaitalicalgrphosphorylationstatusdifferentiallyregulatespyocyaninandpyoverdineproduction AT williamlcody italictoggleyespseudomonasaeruginosaitalicalgrphosphorylationstatusdifferentiallyregulatespyocyaninandpyoverdineproduction AT michaeljschurr italictoggleyespseudomonasaeruginosaitalicalgrphosphorylationstatusdifferentiallyregulatespyocyaninandpyoverdineproduction AT michaellvasil italictoggleyespseudomonasaeruginosaitalicalgrphosphorylationstatusdifferentiallyregulatespyocyaninandpyoverdineproduction AT michaeljschurr italictoggleyespseudomonasaeruginosaitalicalgrphosphorylationstatusdifferentiallyregulatespyocyaninandpyoverdineproduction
_version_	1718427293073801216
spelling	oai:doaj.org-article:a049fc745130465a9bc5434bc9ba9c792021-11-15T15:53:26Z<italic toggle="yes">Pseudomonas aeruginosa</italic> AlgR Phosphorylation Status Differentially Regulates Pyocyanin and Pyoverdine Production10.1128/mBio.02318-172150-7511https://doaj.org/article/a049fc745130465a9bc5434bc9ba9c792018-03-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02318-17https://doaj.org/toc/2150-7511ABSTRACT Pseudomonas aeruginosa employs numerous, complex regulatory elements to control expression of its many virulence systems. The P. aeruginosa AlgZR two-component regulatory system controls the expression of several crucial virulence phenotypes. We recently determined, through transcriptomic profiling of a PAO1 ΔalgR mutant strain compared to wild-type PAO1, that algZR and hemCD are cotranscribed and show differential iron-dependent gene expression. Previous expression profiling was performed in strains without algR and revealed that AlgR acts as either an activator or repressor, depending on the gene. Thus, examination of P. aeruginosa gene expression from cells locked into different AlgR phosphorylation states reveals greater physiological relevance. Therefore, gene expression from strains carrying algR alleles encoding a phosphomimetic (AlgR D54E) or a phosphoablative (AlgR D54N) form were compared by microarray to PAO1. Transcriptome analyses of these strains revealed 25 differentially expressed genes associated with iron siderophore biosynthesis or heme acquisition or production. The PAO1 algR D54N mutant produced lower levels of pyoverdine but increased expression of the small RNAs prrf1 and prrf2 compared to PAO1. In contrast, the algR D54N mutant produced more pyocyanin than wild-type PAO1. On the other hand, the PAO1 algR D54E mutant produced higher levels of pyoverdine, likely due to increased expression of an iron-regulated gene encoding the sigma factor pvdS, but it had decreased pyocyanin production. AlgR specifically bound to the prrf2 and pvdS promoters in vitro. AlgR-dependent pyoverdine production was additionally influenced by carbon source rather than the extracellular iron concentration per se. AlgR phosphorylation effects were also examined in a Drosophila melanogaster feeding, murine acute pneumonia, and punch wound infection models. Abrogation of AlgR phosphorylation attenuated P. aeruginosa virulence in these infection models. These results show that the AlgR phosphorylation state can directly, as well as indirectly, modulate the expression of iron acquisition genes that may ultimately impact the ability of P. aeruginosa to establish and maintain an infection. IMPORTANCE Pyoverdine and pyocyanin production are well-known P. aeruginosa virulence factors that obtain extracellular iron from the environment and from host proteins in different manners. Here, we show that the AlgR phosphorylation state inversely controls pyoverdine and pyocyanin production and that this control is carbon source dependent. P. aeruginosa expressing AlgR D54N, mimicking the constitutively unphosphorylated state, produced more pyocyanin than cells expressing wild-type AlgR. In contrast, a strain expressing an AlgR phosphomimetic (AlgR D54E) produced higher levels of pyoverdine. Pyoverdine production was directly controlled through the prrf2 small regulatory RNA and the pyoverdine sigma factor, PvdS. Abrogating pyoverdine or pyocyanin gene expression has been shown to attenuate virulence in a variety of models. Moreover, the inability to phosphorylate AlgR attenuates virulence in three different models, a Drosophila melanogaster feeding model, a murine acute pneumonia model, and a wound infection model. Interestingly, AlgR-dependent pyoverdine production was responsive to carbon source, indicating that this regulation has additional complexities that merit further study.Alexander S. LittleYuta OkkotsuAlexandria A. ReinhartF. Heath DamronMariette BarbierBrandon BarrettAmanda G. Oglesby-SherrouseJoanna B. GoldbergWilliam L. CodyMichael J. SchurrMichael L. VasilMichael J. SchurrAmerican Society for MicrobiologyarticleAlgRPseudomonas aeruginosairon acquisitionpyocyaninpyoverdinevirulence regulationMicrobiologyQR1-502ENmBio, Vol 9, Iss 1 (2018)

<italic toggle="yes">Pseudomonas aeruginosa</italic> AlgR Phosphorylation Status Differentially Regulates Pyocyanin and Pyoverdine Production

Ejemplares similares